ESTABLISHMENT OF METHOD OF LARGE-SCALE CRYOPRESERVATION OF PORCINE HEPATOCYTES / 解放军医学杂志

ABSTRACT

Objective To establish a method of large-scale cryopreservation of porcine hepatocytes to meet the need of the biological artificial liver. Methods Porcine hepatocytes were isolated from Chinese miniature swines by collagenase disgestion. The cells were suspended in self-made nutrient solution supplemented with 10% DMSO. The hepatocytes (1?10 10~2?10 10) were stored in liquid nitrogen (-196℃) after being treated with two kinds of freezing methods. The cryopreserved hepatocytes were thawed in 39℃ water after one month and cultured. The morphology, viability and the function of thawed hepatocytes were observed. Results The cell viability of cryobreserved hepatocytes after both freezing methods were high. However, synthesis of urea nitrogen and glucose was higher in stepwise temperature lowering group compared with programmed temperature lowering group. Conclusion The results indicate that the cryopreservation method is feasible and simple, large amount porcine hepatocytes can be cryopreserved.